Using Omics to Understand the Effects of Drug Abuse on HIV Latent Reservoir

项目总结/摘要 这项研究的目的是促进对可卡因滥用（CA）与HIV潜伏感染之间的生物学联系的理解。 水库（蓄水池）。我们假设基因调控是一个主要环节。我们提出了一种新的全基因组 RNA表达（RNAexp）研究，以发现从CA到增加的mRNA的基因调控途径， 通过DNA甲基化（DNA m）和介导分析来评估指定途径的联系。 随着联合抗逆转录病毒疗法（cART）和减少艾滋病毒的公共卫生战略的成功， 由于艾滋病的发病率很高，发达国家的艾滋病毒负担现在大部分是一种慢性疾病，包括吸毒者， 用户.作为一种慢性疾病，寻求艾滋病毒治愈方法取决于了解和消除艾滋病毒感染。 (i.e.,有复制能力但沉默的HIV前病毒整合到宿主细胞的DNA中， 停止cART）。可卡因是艾滋病毒阳性者中最常滥用的非法药物之一。 CA与加速HIV进展相关，即使使用cART也会持续存在。作为佐证，我们还 观察到在坚持cART的CA中HIV病毒载量（VL）的下降比坚持非使用者慢。与VL 由于CA与生物量有关，因此了解CA对生物量的影响是至关重要的。 我们专注于基因调控，因为它是CA和CA之间相互作用的生物学连接点。 - 是的很明显，CA和cART都影响基因调控，正常基因调控的破坏是 对HIV病毒复制和传染性很重要，而基因调控在其中起着重要作用 建立和重新激活网络。据我们所知，目前还没有研究量化 通过药物使用整体或CA具体，也没有研究的生物学途径连接CA到ESTA。因此我们 提出以下目标： ·目标1：量化风险并确定与CA相关的差异。 目标2：确定CA对HIV+个体中基因调控的影响。 ·目的3：测试CA差异调节的基因作为CA和ESTA之间的生物介质。 这项研究是重要的和创新的（例如，第一次估计CA中的DNA，第一次全基因组研究， CA对CD 4 T细胞基因调控的影响，作为CA和CD 4 T细胞之间介导者的基因调控的第一次测试 ）。我们的顺序设计，高风险/高回报的建议，适合这个R61/R33机制： 确定在R61期间CA和相关差异基因表达的显著差异 阶段，并评估作为R33期介质的基因调控。这项研究可能会产生 对艾滋病毒作为CA中慢性疾病的关键要素的重要新见解，为 针对CA的独特特征，这些特征增加了感染率，并使HIV治疗在该人群中更具挑战性。

PROJECT SUMMARY/ABSTRACT Goals of this study are to advance understanding of the biology linking cocaine abuse (CA) to HIV latent reservoir (HLR). We hypothesize that gene regulation is a primary link. We propose a novel genome-wide RNA expression (RNAexp) study to discover gene regulatory pathways from CA to increased HLR, followed by DNA methylation (DNAm) and mediation analyses to assess linkages for nominated pathways. With the success of combination antiretroviral therapy (cART) and public health strategies to reduce HIV incidence, much of the HIV burden in developed countries is now as a chronic disease, including among drug users. As a chronic disease, the search for an HIV cure is dependent on understanding and eliminating HLR (i.e., the replication competent but silenced HIV provirus integrated into host cells' DNA, which reactivates with cessation of cART). Cocaine is one of the most frequently abused illicit drugs among HIV+ individuals. CA is associated with accelerated HIV progression, which persists even with cART. Corroboratively, we also observe slower decline in HIV viral load (VL) among cART adherent CAs than adherent nonusers. With VL being associated with HLR quantity, understanding the biological effects of CA on HLR are critical. We focus on gene regulation because it is a biological connecting point for the interplay between CA and HLR. It is clear that both CA and cART affect gene regulation, that disruption of normal gene regulation is important for HIV viral replication and infectivity, and that gene regulation plays an important role in establishing and reactivating HLR. To our knowledge, there are no studies quantifying the likely differences in HLR by drug use overall or by CA specifically, nor studies of biological pathways linking CA to HLR. Thus, we propose the following aims: • Aim 1: Quantify the HLR and determine differences associated with CA. • Aim 2: Determine the effects of CA on gene regulation among HIV+ individuals. • Aim 3: Test CA differentially regulated genes as biological mediators between CA and HLR. This study is significant and innovative (e.g., first estimation of HLR among CAs, first genome-wide study of CA effects on gene regulation in CD4 T cells, first test of gene regulation as a mediator between CA and HLR). Our sequentially designed, high-risk / high-reward proposal, fits well with this R61/R33 mechanism: determining significant differences in HLR by CA and related differential gene expression during the R61 phase, and assessing gene regulation as mediators in the R33 phase. This study is likely to produce important new insights into key elements of HIV as a chronic disease among CAs, providing a basis for targeting unique features of CA that increase HLR and make an HIV cure more challenging in this population.